|
|
| Line 182: |
Line 182: |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br><br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 213: |
Line 213: |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| - | | + | |
| | </div> | | </div> |
| | </div> | | </div> |
| Line 243: |
Line 242: |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 274: |
Line 273: |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 305: |
Line 304: |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | </div> | | </div> |
| Line 352: |
Line 351: |
| | <a href="#" class="head">Week 1</a> | | <a href="#" class="head">Week 1</a> |
| | <div class="content" align="left" style="display: none;"> | | <div class="content" align="left" style="display: none;"> |
| - | <br> <b> July 2 </b> <br> | + | <br> <b> July xx </b> <br> |
| - | Wet lab <br> | + | Wet lab<br> |
| - | · Digestion of pEGFP-N1 by Ase1 and BamH1 for PPAR-alpha promoter; , Xba1 and BamH1 for FABP1 promoter <br> | + | · <br> |
| - | · LB-Chloramphenicol plates (the previous ones were found contaminated) <br> | + | · <br> |
| - | · Inoculation of pEGFP-N1 for plasmid extraction <br> | + | · <br> |
| - | · Transformation of P<i>fadBA</i> due chloramphenicol plates contamination <br> | + | · <br> |
| | | | |
| - | <br> <b> July 3 </b> <br> | + | <br> <b> July xx </b> <br> |
| - | · Extraction of pEGFP-N1 plasmids by miniprep <br>
| + | |
| - | · PCR for PPAR-alpha promoter amplification <br>
| + | |
| - | · Digestion of pEGFP-N1 by Ase1 and BamH1 for PPAR-alpha promoter; , Xba1 and BamH1 for FABP1 promoter <br>
| + | |
| - | | + | |
| - | <br> <b> July 4</b> <br>
| + | |
| | Wet lab<br> | | Wet lab<br> |
| - | · Inoculation of BBa_K817002 P<i>fadBA</i><br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter and FABP1 promoter cloning from gDNA<br> | + | · <br> |
| - | · Ran gel for digestion of pEGFP-N1 by Ase1 and BamH1 for PPAR-alpha promoter; Xba1 and BamH1 for FABP1 promoter; Ase1 and Xhol1 for GRP78<br> | + | · <br> |
| | + | · <br> |
| | | | |
| - | <br> <b> July 5</b> <br> | + | <br> <b> July xx</b> <br> |
| | + | Wet lab<br> |
| | + | · <br> |
| | + | · <br> |
| | + | · <br> |
| | + | · <br> |
| | + | <br> <b> July xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Extraction of BBa_K817002 (P<i>fadBA</i>) <br> | + | · <br> |
| - | · Inoculation of pEGFP-N1<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter and FABP1 promoter cloning from gDNA<br> | + | · <br> |
| - | · Gel check, 0.8% gel for previously gDNA extraction<br> | + | · <br> |
| - | · Ran gel for PCR check of PPAR-alpha promoter and FABP1 promoter<br>
| + | |
| - | · Poured new LB-Kanamycin plates<br>
| + | |
| - | · Transformation of <a href="http://parts.igem.org/Part:BBa_J176171">BBa_J176171</a><br>
| + | |
| - | Dry lab<br>
| + | |
| - | · Discussion about re-design constructions, possible change for BBa_J176171 as mammalian expression vector<br>
| + | |
| - | · Primer redesign for PPAR-alpha promoter and FABP1 promoter<br><br>
| + | |
| | | | |
| | | | |
| Line 390: |
Line 385: |
| | <a href="#" class="head">Week 2</a> | | <a href="#" class="head">Week 2</a> |
| | <div class="content" align="left" style="display: none;"> | | <div class="content" align="left" style="display: none;"> |
| - | <br> <b> July 8</b> <br> | + | <br> <b> July xx </b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · gDNA extraction from HepG2 cells<br> | + | · <br> |
| - | · Restriction of BBa_K817002 P<i>fadBA</i><br> | + | · <br> |
| - | · Gel check for gDNA extraction<br> | + | · <br> |
| - | · Inoculation of BBa_J176171, BBa_K817002 (P<i>fadBA</i>), pEGFP-N1 for plasmid extraction<br> | + | · <br> |
| - | · New primers for PPAR-alpha promoter and FABP1 promoter arrival<br>
| + | |
| - | · Ran gel for BBa_K817002 P<i>fadBA</i> restriction check, gel extraction and purification<br>
| + | |
| | | | |
| - | <br> <b> July 9</b> <br> | + | <br> <b> July xx </b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Extraction of BBa_J176171, BBa_K817002 P<i>fadBA</i> and pEGFP-N1 by miniprep<br> | + | · <br> |
| - | · Restriction of pEGFP-N1 Restriction digestion by Ase1 and BamH1 for PPAR-alpha promoter; Ase1 and Xho1 for GRP78, Xba1 and BamH1 for FABP1 promoter, Pst1 and Not1; followed by gel check, extraction and purification<br> | + | · <br> |
| - | · Restriction of BBa_J176171 for P<i>fadBA</i> Ase1 and BamH1; FABP1 promoter by Xba1 and Not1<br> | + | · <br> |
| - | · Ran gel for pEGFP-N1 and BBa_J176171 restriction products<br> | + | · <br> |
| - | · PCR PPAR-alpha promoter and FABP1 promoter cloning from gDNA<br>
| + | |
| - | · <a href="http://parts.igem.org/Part:BBa_J52034">BBa_J52034</a> (<i>fad</i>R) transformation<br>
| + | |
| | | | |
| - | <br> <b> July 10</b> <br> | + | <br> <b> July xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Ran gel for PCR check of PPAR-alpha promoter and FAPB1 promoter<br> | + | · <br> |
| - | · PCR for FABP1 promoter, PCR replication<br> | + | · <br> |
| - | · BBa_J176171 vector dephosphorylation by antartic phosphatase<br> | + | · <br> |
| - | · Ligation of BBa_K817002 and P<i>fadBA</i> and BBa_J176171 <br> | + | · <br> |
| - | · BBa_J52034 (<i>fad</i>R) inoculation<br>
| + | <br> <b> July xx</b> <br> |
| - | · Digestion for BBa_K817002 P<i>fadBA</i> extraction<br>
| + | |
| - | Gel check for BBa_K817002 P<i>fadBA</i> extraction<br>
| + | |
| - | | + | |
| - | <br> <b> July 11</b> <br>
| + | |
| - | Wet lab<br>
| + | |
| - | · Gel purification for BBa_K817002 P<i>fadBA</i> extraction<br>
| + | |
| - | · Gel check for FABP1 promoter PCR product<br>
| + | |
| - | · PCR clean-up<br>
| + | |
| - | · BBa_J52034 (<i>fad</i>R) miniprep<br>
| + | |
| - | · BBa_J52034 restriction by Pst1 HF and Not1 HF<br>
| + | |
| - | · PCR for PPAR-alpha promoter<br>
| + | |
| - | Dry lab<br>
| + | |
| - | Re-design constructions, now using BBa_J176171 as mammalian expression vector for all related constructions<br>
| + | |
| - | | + | |
| - | <br> <b> July 12</b> <br> | + | |
| | Wet lab<br> | | Wet lab<br> |
| - | · Digestion of FABP1 promoter PCR product<br>
| + | · <br> |
| - | · Ran gel for PCR check of PPAR-alpha promoter<br> | + | · <br> |
| - | · Digest pEGFP-n1 for BBa_J52034 <i>fad</i>R<br> | + | · <br> |
| - | · Ran gel for pEGFP-n1 for BBa_J52034 <i>fad</i>R followed by gel purification<br> | + | · <br> |
| - | · BBa_J176171 vector dephosphorylation by Antarctic phosphatase for FABP1 promoter and P<i>fadBA</i><br> | + | |
| - | Dry Lab<br>
| + | |
| - | Primers design for pCMV cloning for <i>fad</i>R expression<br><br>
| + | |
| | </div> | | </div> |
| | </div> | | </div> |
| Line 444: |
Line 417: |
| | <a href="#" class="head">Week 3</a> | | <a href="#" class="head">Week 3</a> |
| | <div class="content" align="left" style="display: none;"> | | <div class="content" align="left" style="display: none;"> |
| - | <br><b>July 15</b> <br> | + | <br> <b> July xx </b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Ligation of FABP1 promoter, EGFP, and BBa_J176171, using 3 pieces ligation<br> | + | · <br> |
| - | · Transformation of FABP1 promoter, EGFP, and BBa_J176171 ligation<br> | + | · <br> |
| - | · FABP1 promoter+EGFP +BBa_J176171 ligation restriction check<br> | + | · <br> |
| | + | · <br> |
| | | | |
| - | <br><b>July 16</b> <br> | + | <br> <b> July xx </b> <br> |
| - | Wet Lab<br>
| + | |
| - | · Miniprep for full construct of FABP1 promoter and pEGFP-N1<br>
| + | |
| - | · <i>fad</i>R and pEGFP-N1 Backbone parts ligation <br>
| + | |
| - | · Plasmids extraction for pCMV cloning for <i>fad</i>R<br>
| + | |
| - | · BBa_K817002 P<i>fadBA</i> promoter extraction by EcoR1 and Pst1 HF<br>
| + | |
| - | · BBa_J52034 restriction by EcoR1 and Pst1 HF<br>
| + | |
| - | · Inoculations for full construct of FABP1 promoter and pEGFP-N1<br>
| + | |
| - | · Streak colonies containing the right construct for FABP1 promoter<br>
| + | |
| - | | + | |
| - | <br><b>July 17</b> <br>
| + | |
| | Wet lab<br> | | Wet lab<br> |
| - | · Plasmid extraction for FABP1 promoter+EGFP+BBa_J176171 by miniprep<br> | + | · <br> |
| - | · Repeat <i>fad</i>R and P<i>fadBA</i> constructs<br> | + | · <br> |
| - | · Digest pEGFP-n1 and BBa-J176171 for <i>fad</i>R<br> | + | · <br> |
| - | · Digestion for BBa_K817002 (P<i>fadBA</i>) extraction<br> | + | · <br> |
| - | · Gel check and gel extraction for BBa_J176171 for <i>fad</i>R and BBa_K817002 (P<i>fadBA</i>) <br>
| + | |
| | | | |
| - | <br><b>July 18</b> <br> | + | <br> <b> July xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Gel purification for all digested products<br> | + | · <br> |
| - | · FABP1 promoter digestion check for whole construct prior transfection<br> | + | · <br> |
| - | · P<i>fadBA</i> vector de phosphorylation and ligation with EGFP and BBa_J176171<br> | + | · <br> |
| - | · Transformation of P<i>fadBA</i>+EGFP+BBa_J176171<br> | + | · <br> |
| - | | + | <br> <b> July xx</b> <br> |
| - | <br><b>July 19</b> <br> | + | |
| | Wet lab<br> | | Wet lab<br> |
| - | · FABP1 promoter preparation for transfection<br> | + | · <br> |
| - | · Ligation of P<i>fadBA</i> and BBa_J176171 followed by transformation<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter cloning<br> | + | · <br> |
| - | Dry lab<br>
| + | · <br> |
| - | · Design for characterization of the transfected cells with FABP1 construct<br> | + | |
| - | · Review for Multiple Sites Mutagenesis<br>
| + | |
| - | · Ordered primers for pCMV cloning from pEGFP-N1<br><br>
| + | |
| | | | |
| | </div> | | </div> |
| Line 491: |
Line 450: |
| | <a href="#" class="head">Week 4</a> | | <a href="#" class="head">Week 4</a> |
| | <div class="content" align="left" style="display: none;"> | | <div class="content" align="left" style="display: none;"> |
| - | <br><b>July 22</b> <br> | + | <br> <b> July xx </b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · FABP1 construct given for characterization and transfection<br> | + | · <br> |
| - | · Further colony screening for FABP1 construct, inoculations<br> | + | · <br> |
| - | · Inoculation of P<i>fadBA</i> and BBa_J176171<br> | + | · <br> |
| - | · Ran gel for PPAR-alpha promoter PCR products<br> | + | · <br> |
| | | | |
| - | <br><b>July 23</b> <br> | + | <br> <b> July xx </b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Primers arrival for pCMV cloning<br> | + | · <br> |
| - | · PCR for pCMV cloning<br> | + | · <br> |
| - | · Digestion and gel for construction check for FABP1 promoter<br> | + | · <br> |
| - | · P<i>fadBA</i> gel extraction and purification, followed by vector dephosphorylation BBa_J176171 and ligation, then transformation<br> | + | · <br> |
| - | · Digestion of pEGFP-N1 for <i>fad</i>R, followed by gel check and extraction<br>
| + | |
| | | | |
| - | <br><b>July 24</b> <br> | + | <br> <b> July xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · DNA purification from gel extraction for digested pEGFP-N1<br> | + | · <br> |
| - | · Multiple sites mutagenesis for illegal restriction sites for FABP1 promoter<br> | + | · <br> |
| - | · Ran gel for pCMV cloning from pEGFP-N1<br> | + | · <br> |
| - | · PCR clean up for pCMV cloning from pEGFP-N1<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter with new primers using Taq polymerase<br>
| + | <br> <b> July xx</b> <br> |
| - | · Transformation of mutagenesis products<br>
| + | |
| - | · Inoculation of P<i>fadBA</i> ligation colonies<br>
| + | |
| - | | + | |
| - | <br><b>July 25</b> <br> | + | |
| | Wet lab<br> | | Wet lab<br> |
| - | · Ran gel check PPAR-alpha promoter PCR cloning with new primers using Taq polymerase<br> | + | · <br> |
| - | · Minprep of P<i>fadBA</i>+EGFP+BBa_J176171 ligation colonies, followed by digestion check <br> | + | · <br> |
| - | | + | · <br> |
| - | <br><b>July 26</b> <br>
| + | · <br> |
| - | Wet lab<br>
| + | |
| - | · Multiple sites mutagenesis for illegal restriction sites for FABP1 promoter<br> | + | |
| - | · Ran gel before parental string digestion of mutagenesis products<br> | + | |
| - | · Digestion of pEGFP-N1 by Apa1 and Xba1 followed by gel check<br>
| + | |
| - | · Plasmids preparation according to transfection requirements, construct and controls<br><br>
| + | |
| - | | + | |
| | </div> | | </div> |
| | </div> | | </div> |
| Line 534: |
Line 482: |
| | <a href="#" class="head">Week 5</a> | | <a href="#" class="head">Week 5</a> |
| | <div class="content" align="left" style="display: none;"> | | <div class="content" align="left" style="display: none;"> |
| - | <br><b>July 30</b> <br> | + | <br> <b> July xx </b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · PPAR-alpha promoter PCR cloning with new primers using Vent polymerase<br> | + | · <br> |
| - | · Restriction check for pEGFP-N1, using EcoR1, Xba1, Spe1 and Pst1<br> | + | · <br> |
| - | · BBa_J176171 vector de phosphorylation, followed by ligation with EGFP using T4 Ligase, then transformation into E. Coli strain DH10b<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter with new primers using Vent polymerase<br> | + | · <br> |
| - | Dry lab<br>
| + | |
| - | · Discussion and re assessment of constructs related to pEGFP-N1<br>
| + | |
| | | | |
| | + | <br> <b> July xx </b> <br> |
| | + | Wet lab<br> |
| | + | · <br> |
| | + | · <br> |
| | + | · <br> |
| | + | · <br> |
| | | | |
| - | <br><b>July 31</b> <br> | + | <br> <b> July xx</b> <br> |
| - | · Ran gel check for PPAR-alpha promoter PCR cloning with new primers using Vent polymerase<br> | + | Wet lab<br> |
| - | · PCR for pCMV cloning from pEGFP-N1, followed by gel check and PCR clean up<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter with reference primers using Vent polymerase<br> | + | · <br> |
| - | · Ran gel check for PPAR-alpha promoter PCR cloning with reference primers using Vent polymerase<br><br> | + | · <br> |
| | + | · <br> |
| | + | <br> <b> July xx</b> <br> |
| | + | Wet lab<br> |
| | + | · <br> |
| | + | · <br> |
| | + | · <br> |
| | + | · <br> |
| | | | |
| | </div> | | </div> |
| Line 573: |
Line 532: |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 604: |
Line 563: |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | | | |
| | </div> | | </div> |
| Line 634: |
Line 594: |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | | | |
| | | | |
| Line 665: |
Line 626: |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 696: |
Line 657: |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 743: |
Line 704: |
| | <a href="#" class="head">Week 1</a> | | <a href="#" class="head">Week 1</a> |
| | <div class="content" align="left" style="display: none;"> | | <div class="content" align="left" style="display: none;"> |
| - | <br><b>August 2</b> <br> | + | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 782: |
Line 747: |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| - | | + | |
| | </div> | | </div> |
| | </div> | | </div> |
| Line 818: |
Line 787: |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 855: |
Line 829: |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 892: |
Line 870: |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 999: |
Line 982: |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br><br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 1,030: |
Line 1,013: |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| - | | + | |
| | </div> | | </div> |
| | </div> | | </div> |
| Line 1,060: |
Line 1,042: |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 1,091: |
Line 1,073: |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 1,122: |
Line 1,104: |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>June xx</b> <br> | | <br><b>June xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | </div> | | </div> |
| Line 1,169: |
Line 1,151: |
| | <a href="#" class="head">Week 1</a> | | <a href="#" class="head">Week 1</a> |
| | <div class="content" align="left" style="display: none;"> | | <div class="content" align="left" style="display: none;"> |
| - | <br> <b> July 2 </b> <br> | + | <br> <b> July xx </b> <br> |
| - | Wet lab <br> | + | Wet lab<br> |
| - | · Digestion of pEGFP-N1 by Ase1 and BamH1 for PPAR-alpha promoter; , Xba1 and BamH1 for FABP1 promoter <br> | + | · <br> |
| - | · LB-Chloramphenicol plates (the previous ones were found contaminated) <br> | + | · <br> |
| - | · Inoculation of pEGFP-N1 for plasmid extraction <br> | + | · <br> |
| - | · Transformation of P<i>fadBA</i> due chloramphenicol plates contamination <br> | + | · <br> |
| | | | |
| - | <br> <b> July 3 </b> <br> | + | <br> <b> July xx </b> <br> |
| - | · Extraction of pEGFP-N1 plasmids by miniprep <br>
| + | |
| - | · PCR for PPAR-alpha promoter amplification <br>
| + | |
| - | · Digestion of pEGFP-N1 by Ase1 and BamH1 for PPAR-alpha promoter; , Xba1 and BamH1 for FABP1 promoter <br>
| + | |
| - | | + | |
| - | <br> <b> July 4</b> <br>
| + | |
| | Wet lab<br> | | Wet lab<br> |
| - | · Inoculation of BBa_K817002 P<i>fadBA</i><br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter and FABP1 promoter cloning from gDNA<br> | + | · <br> |
| - | · Ran gel for digestion of pEGFP-N1 by Ase1 and BamH1 for PPAR-alpha promoter; Xba1 and BamH1 for FABP1 promoter; Ase1 and Xhol1 for GRP78<br> | + | · <br> |
| | + | · <br> |
| | | | |
| - | <br> <b> July 5</b> <br> | + | <br> <b> July xx</b> <br> |
| | + | Wet lab<br> |
| | + | · <br> |
| | + | · <br> |
| | + | · <br> |
| | + | · <br> |
| | + | <br> <b> July xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Extraction of BBa_K817002 (P<i>fadBA</i>) <br> | + | · <br> |
| - | · Inoculation of pEGFP-N1<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter and FABP1 promoter cloning from gDNA<br> | + | · <br> |
| - | · Gel check, 0.8% gel for previously gDNA extraction<br> | + | · <br> |
| - | · Ran gel for PCR check of PPAR-alpha promoter and FABP1 promoter<br>
| + | |
| - | · Poured new LB-Kanamycin plates<br>
| + | |
| - | · Transformation of <a href="http://parts.igem.org/Part:BBa_J176171">BBa_J176171</a><br>
| + | |
| - | Dry lab<br>
| + | |
| - | · Discussion about re-design constructions, possible change for BBa_J176171 as mammalian expression vector<br>
| + | |
| - | · Primer redesign for PPAR-alpha promoter and FABP1 promoter<br><br>
| + | |
| | | | |
| | | | |
| Line 1,207: |
Line 1,185: |
| | <a href="#" class="head">Week 2</a> | | <a href="#" class="head">Week 2</a> |
| | <div class="content" align="left" style="display: none;"> | | <div class="content" align="left" style="display: none;"> |
| - | <br> <b> July 8</b> <br> | + | <br> <b> July xx </b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · gDNA extraction from HepG2 cells<br> | + | · <br> |
| - | · Restriction of BBa_K817002 P<i>fadBA</i><br> | + | · <br> |
| - | · Gel check for gDNA extraction<br> | + | · <br> |
| - | · Inoculation of BBa_J176171, BBa_K817002 (P<i>fadBA</i>), pEGFP-N1 for plasmid extraction<br> | + | · <br> |
| - | · New primers for PPAR-alpha promoter and FABP1 promoter arrival<br>
| + | |
| - | · Ran gel for BBa_K817002 P<i>fadBA</i> restriction check, gel extraction and purification<br>
| + | |
| | | | |
| - | <br> <b> July 9</b> <br> | + | <br> <b> July xx </b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Extraction of BBa_J176171, BBa_K817002 P<i>fadBA</i> and pEGFP-N1 by miniprep<br> | + | · <br> |
| - | · Restriction of pEGFP-N1 Restriction digestion by Ase1 and BamH1 for PPAR-alpha promoter; Ase1 and Xho1 for GRP78, Xba1 and BamH1 for FABP1 promoter, Pst1 and Not1; followed by gel check, extraction and purification<br> | + | · <br> |
| - | · Restriction of BBa_J176171 for P<i>fadBA</i> Ase1 and BamH1; FABP1 promoter by Xba1 and Not1<br> | + | · <br> |
| - | · Ran gel for pEGFP-N1 and BBa_J176171 restriction products<br> | + | · <br> |
| - | · PCR PPAR-alpha promoter and FABP1 promoter cloning from gDNA<br>
| + | |
| - | · <a href="http://parts.igem.org/Part:BBa_J52034">BBa_J52034</a> (<i>fad</i>R) transformation<br>
| + | |
| | | | |
| - | <br> <b> July 10</b> <br> | + | <br> <b> July xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Ran gel for PCR check of PPAR-alpha promoter and FAPB1 promoter<br> | + | · <br> |
| - | · PCR for FABP1 promoter, PCR replication<br> | + | · <br> |
| - | · BBa_J176171 vector dephosphorylation by antartic phosphatase<br> | + | · <br> |
| - | · Ligation of BBa_K817002 and P<i>fadBA</i> and BBa_J176171 <br> | + | · <br> |
| - | · BBa_J52034 (<i>fad</i>R) inoculation<br>
| + | <br> <b> July xx</b> <br> |
| - | · Digestion for BBa_K817002 P<i>fadBA</i> extraction<br>
| + | |
| - | Gel check for BBa_K817002 P<i>fadBA</i> extraction<br>
| + | |
| - | | + | |
| - | <br> <b> July 11</b> <br>
| + | |
| - | Wet lab<br>
| + | |
| - | · Gel purification for BBa_K817002 P<i>fadBA</i> extraction<br>
| + | |
| - | · Gel check for FABP1 promoter PCR product<br>
| + | |
| - | · PCR clean-up<br>
| + | |
| - | · BBa_J52034 (<i>fad</i>R) miniprep<br>
| + | |
| - | · BBa_J52034 restriction by Pst1 HF and Not1 HF<br>
| + | |
| - | · PCR for PPAR-alpha promoter<br>
| + | |
| - | Dry lab<br>
| + | |
| - | Re-design constructions, now using BBa_J176171 as mammalian expression vector for all related constructions<br>
| + | |
| - | | + | |
| - | <br> <b> July 12</b> <br> | + | |
| | Wet lab<br> | | Wet lab<br> |
| - | · Digestion of FABP1 promoter PCR product<br>
| + | · <br> |
| - | · Ran gel for PCR check of PPAR-alpha promoter<br> | + | · <br> |
| - | · Digest pEGFP-n1 for BBa_J52034 <i>fad</i>R<br> | + | · <br> |
| - | · Ran gel for pEGFP-n1 for BBa_J52034 <i>fad</i>R followed by gel purification<br> | + | · <br> |
| - | · BBa_J176171 vector dephosphorylation by Antarctic phosphatase for FABP1 promoter and P<i>fadBA</i><br> | + | |
| - | Dry Lab<br>
| + | |
| - | Primers design for pCMV cloning for <i>fad</i>R expression<br><br>
| + | |
| | </div> | | </div> |
| | </div> | | </div> |
| Line 1,261: |
Line 1,217: |
| | <a href="#" class="head">Week 3</a> | | <a href="#" class="head">Week 3</a> |
| | <div class="content" align="left" style="display: none;"> | | <div class="content" align="left" style="display: none;"> |
| - | <br><b>July 15</b> <br> | + | <br> <b> July xx </b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Ligation of FABP1 promoter, EGFP, and BBa_J176171, using 3 pieces ligation<br> | + | · <br> |
| - | · Transformation of FABP1 promoter, EGFP, and BBa_J176171 ligation<br> | + | · <br> |
| - | · FABP1 promoter+EGFP +BBa_J176171 ligation restriction check<br> | + | · <br> |
| | + | · <br> |
| | | | |
| - | <br><b>July 16</b> <br> | + | <br> <b> July xx </b> <br> |
| - | Wet Lab<br>
| + | |
| - | · Miniprep for full construct of FABP1 promoter and pEGFP-N1<br>
| + | |
| - | · <i>fad</i>R and pEGFP-N1 Backbone parts ligation <br>
| + | |
| - | · Plasmids extraction for pCMV cloning for <i>fad</i>R<br>
| + | |
| - | · BBa_K817002 P<i>fadBA</i> promoter extraction by EcoR1 and Pst1 HF<br>
| + | |
| - | · BBa_J52034 restriction by EcoR1 and Pst1 HF<br>
| + | |
| - | · Inoculations for full construct of FABP1 promoter and pEGFP-N1<br>
| + | |
| - | · Streak colonies containing the right construct for FABP1 promoter<br>
| + | |
| - | | + | |
| - | <br><b>July 17</b> <br>
| + | |
| | Wet lab<br> | | Wet lab<br> |
| - | · Plasmid extraction for FABP1 promoter+EGFP+BBa_J176171 by miniprep<br> | + | · <br> |
| - | · Repeat <i>fad</i>R and P<i>fadBA</i> constructs<br> | + | · <br> |
| - | · Digest pEGFP-n1 and BBa-J176171 for <i>fad</i>R<br> | + | · <br> |
| - | · Digestion for BBa_K817002 (P<i>fadBA</i>) extraction<br> | + | · <br> |
| - | · Gel check and gel extraction for BBa_J176171 for <i>fad</i>R and BBa_K817002 (P<i>fadBA</i>) <br>
| + | |
| | | | |
| - | <br><b>July 18</b> <br> | + | <br> <b> July xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Gel purification for all digested products<br> | + | · <br> |
| - | · FABP1 promoter digestion check for whole construct prior transfection<br> | + | · <br> |
| - | · P<i>fadBA</i> vector de phosphorylation and ligation with EGFP and BBa_J176171<br> | + | · <br> |
| - | · Transformation of P<i>fadBA</i>+EGFP+BBa_J176171<br> | + | · <br> |
| - | | + | <br> <b> July xx</b> <br> |
| - | <br><b>July 19</b> <br> | + | |
| | Wet lab<br> | | Wet lab<br> |
| - | · FABP1 promoter preparation for transfection<br> | + | · <br> |
| - | · Ligation of P<i>fadBA</i> and BBa_J176171 followed by transformation<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter cloning<br> | + | · <br> |
| - | Dry lab<br>
| + | · <br> |
| - | · Design for characterization of the transfected cells with FABP1 construct<br> | + | |
| - | · Review for Multiple Sites Mutagenesis<br>
| + | |
| - | · Ordered primers for pCMV cloning from pEGFP-N1<br><br>
| + | |
| | | | |
| | </div> | | </div> |
| Line 1,308: |
Line 1,250: |
| | <a href="#" class="head">Week 4</a> | | <a href="#" class="head">Week 4</a> |
| | <div class="content" align="left" style="display: none;"> | | <div class="content" align="left" style="display: none;"> |
| - | <br><b>July 22</b> <br> | + | <br> <b> July xx </b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · FABP1 construct given for characterization and transfection<br> | + | · <br> |
| - | · Further colony screening for FABP1 construct, inoculations<br> | + | · <br> |
| - | · Inoculation of P<i>fadBA</i> and BBa_J176171<br> | + | · <br> |
| - | · Ran gel for PPAR-alpha promoter PCR products<br> | + | · <br> |
| | | | |
| - | <br><b>July 23</b> <br> | + | <br> <b> July xx </b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Primers arrival for pCMV cloning<br> | + | · <br> |
| - | · PCR for pCMV cloning<br> | + | · <br> |
| - | · Digestion and gel for construction check for FABP1 promoter<br> | + | · <br> |
| - | · P<i>fadBA</i> gel extraction and purification, followed by vector dephosphorylation BBa_J176171 and ligation, then transformation<br> | + | · <br> |
| - | · Digestion of pEGFP-N1 for <i>fad</i>R, followed by gel check and extraction<br>
| + | |
| | | | |
| - | <br><b>July 24</b> <br> | + | <br> <b> July xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · DNA purification from gel extraction for digested pEGFP-N1<br> | + | · <br> |
| - | · Multiple sites mutagenesis for illegal restriction sites for FABP1 promoter<br> | + | · <br> |
| - | · Ran gel for pCMV cloning from pEGFP-N1<br> | + | · <br> |
| - | · PCR clean up for pCMV cloning from pEGFP-N1<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter with new primers using Taq polymerase<br>
| + | <br> <b> July xx</b> <br> |
| - | · Transformation of mutagenesis products<br>
| + | |
| - | · Inoculation of P<i>fadBA</i> ligation colonies<br>
| + | |
| - | | + | |
| - | <br><b>July 25</b> <br> | + | |
| | Wet lab<br> | | Wet lab<br> |
| - | · Ran gel check PPAR-alpha promoter PCR cloning with new primers using Taq polymerase<br> | + | · <br> |
| - | · Minprep of P<i>fadBA</i>+EGFP+BBa_J176171 ligation colonies, followed by digestion check <br> | + | · <br> |
| - | | + | · <br> |
| - | <br><b>July 26</b> <br>
| + | · <br> |
| - | Wet lab<br>
| + | |
| - | · Multiple sites mutagenesis for illegal restriction sites for FABP1 promoter<br> | + | |
| - | · Ran gel before parental string digestion of mutagenesis products<br> | + | |
| - | · Digestion of pEGFP-N1 by Apa1 and Xba1 followed by gel check<br>
| + | |
| - | · Plasmids preparation according to transfection requirements, construct and controls<br><br>
| + | |
| - | | + | |
| | </div> | | </div> |
| | </div> | | </div> |
| Line 1,351: |
Line 1,282: |
| | <a href="#" class="head">Week 5</a> | | <a href="#" class="head">Week 5</a> |
| | <div class="content" align="left" style="display: none;"> | | <div class="content" align="left" style="display: none;"> |
| - | <br><b>July 30</b> <br> | + | <br> <b> July xx </b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · PPAR-alpha promoter PCR cloning with new primers using Vent polymerase<br> | + | · <br> |
| - | · Restriction check for pEGFP-N1, using EcoR1, Xba1, Spe1 and Pst1<br> | + | · <br> |
| - | · BBa_J176171 vector de phosphorylation, followed by ligation with EGFP using T4 Ligase, then transformation into E. Coli strain DH10b<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter with new primers using Vent polymerase<br> | + | · <br> |
| - | Dry lab<br>
| + | |
| - | · Discussion and re assessment of constructs related to pEGFP-N1<br>
| + | |
| | | | |
| | + | <br> <b> July xx </b> <br> |
| | + | Wet lab<br> |
| | + | · <br> |
| | + | · <br> |
| | + | · <br> |
| | + | · <br> |
| | | | |
| - | <br><b>July 31</b> <br> | + | <br> <b> July xx</b> <br> |
| - | · Ran gel check for PPAR-alpha promoter PCR cloning with new primers using Vent polymerase<br> | + | Wet lab<br> |
| - | · PCR for pCMV cloning from pEGFP-N1, followed by gel check and PCR clean up<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter with reference primers using Vent polymerase<br> | + | · <br> |
| - | · Ran gel check for PPAR-alpha promoter PCR cloning with reference primers using Vent polymerase<br><br> | + | · <br> |
| | + | · <br> |
| | + | <br> <b> July xx</b> <br> |
| | + | Wet lab<br> |
| | + | · <br> |
| | + | · <br> |
| | + | · <br> |
| | + | · <br> |
| | | | |
| | </div> | | </div> |
| Line 1,390: |
Line 1,332: |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 1,421: |
Line 1,363: |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | | | |
| | </div> | | </div> |
| Line 1,451: |
Line 1,394: |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | | | |
| | | | |
| Line 1,482: |
Line 1,426: |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 1,513: |
Line 1,457: |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | <br><b>Aug xx</b> <br> | | <br><b>Aug xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 1,560: |
Line 1,504: |
| | <a href="#" class="head">Week 1</a> | | <a href="#" class="head">Week 1</a> |
| | <div class="content" align="left" style="display: none;"> | | <div class="content" align="left" style="display: none;"> |
| - | <br><b>August 2</b> <br> | + | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 1,599: |
Line 1,547: |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| - | | + | |
| | </div> | | </div> |
| | </div> | | </div> |
| Line 1,635: |
Line 1,587: |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 1,672: |
Line 1,629: |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
| Line 1,709: |
Line 1,670: |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | + | |
| | <br><b>Sept xx</b> <br> | | <br><b>Sept xx</b> <br> |
| | Wet lab<br> | | Wet lab<br> |
| - | · Restriction check for EGFP+ BBa_J176171 using EcoR1 and Pst1HF<br> | + | · <br> |
| - | · Consensus Kozak sequence extraction from BBa_J176171 by EcoR1 and Xba1, followed by gel check and DNA purification<br> | + | · <br> |
| - | · PCR for PPAR-alpha promoter, using temperature gradient and every available set of primers designed, using Vent polymerase<br> | + | · <br> |
| - | · Genomic DNA extraction from HepG2 cells, Phenol/Chloroform protocol<br><br> | + | · <br> |
| | | | |
| | | | |
"
